ObjectiveTo investigate the effect of Yaoshu Zhuyu Fang on the regulation of the microRNA-17-5P (miR-17-5P)/murine double minute 2 (MDM2)/p53 axis in the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells, and its potential molecular mechanism. MethodsIntervertebral disc annulus fibrosus tissues were obtained from 8-week-old SPF-grade male SD rats, and annulus fibrosus cells were isolated and obtained by enzyme digestion and mechanical dispersion. Annulus fibrosus cells were divided into 6 groups: Group C was the blank control group, in which annulus fibrosus cells were not treated with interleukin-1β (IL-1β) but were cultured in RPMI 1640 complete medium. Group β was the degeneration model group constructed by treating annulus fibrosus cells with 10 ng/mL IL-1β for 24 h. Group β+B was the IL-1β + blank serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% blank serum for 24 h. Group β+W was the IL-1β + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. Group β+I was the IL-1β + miR-17-5P inhibitor group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor. Group β+I+W was the IL-1β + miR-17-5P inhibitor + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor, and finally treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. CCK-8 assay was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the expression levels of miR-17-5P, MDM2 mRNA, and p53 mRNA in cells. Western blotting was used to detect the protein expression levels of MDM2 and p53 in cells. Dual-luciferase reporter system was used to analyze the targeting relationship between miR-17-5P and MDM2. ResultsCompared with Group C, Group β showed a significant decrease in cell survival rate (P<0.001), a significant increase in cell apoptosis rate (P<0.001), significantly increased expression of miR-17-5P, p53 mRNA, and p53 protein (P<0.001), and significantly decreased expression of MDM2 mRNA and protein (P<0.001). Compared with Group β, Group β+W, Group β+I, and Group β+I+W showed significantly increased cell survival rate, significantly decreased apoptosis rate, significantly decreased expression of miR-17-5P, p53 mRNA, and p53 protein, and significantly increased expression of MDM2 mRNA and protein (P<0.001). Moreover, changes in the above indicators were greater in Group β+I+W (P<0.001). Circular RNA Interactome predicted that miR-17-5P had specific binding sites with the 3' untranslated region (3'UTR) of MDM2. Transfection of miR-17-5P mimic significantly reduced the luciferase expression level of co-transfected luciferase reporter plasmid containing wild-type MDM2 3'UTR (P<0.05), but had no significant effect on luciferase expression in cells co-transfected with luciferase reporter plasmid containing mutant MDM2 3'UTR (P>0.05). ConclusionYaoshu Zhuyu Fang down-regulates the expression of miR-17-5P, promotes the synthesis of MDM2 protein, thereby down-regulates p53, promotes proliferation, and inhibits the apoptosis of rat intervertebral disc annulus fibrosus cells.

Objective: To analyze the process of handling a pulmonary tuberculosis(TB) outbreak among senior high school students in a boarding school in Chongqing, as well as to investigate the underlying causes of the outbreak, so as to provide evidence to inform TB prevention and control strategies in school settings. Methods: From November 2023 to April 2024, an epidemiological investigation was conducted into the TB outbreak in a grade 12 class from a boarding high school. Suspected cases were screened using symptom screening, tuberculin skin test (TST), and chest X-ray examinations. Confirmed cases underwent individual epidemiological interviews and sputum culture; Cultured positive mycobacterial strains were subjected to whole genome sequencing after identification as Mycobacterium tuberculosis. Results: A total of 10 active pulmonary TB cases were identified, all from the same class, yielding a student attack rate of 16.67%. Three isolates were culture positive, as well as all strains were of L2 type，and the WGS analysis of the strains suggested a common transmission chain. Excluding the index case, four additional cases were detected through symptom driven health care visits. Notably, 70% of patients presented with "chest tightness and chest pain" symptoms, and 50% had "cough" symptoms,but none were detected during morning health checks or tracking of absences due to illness. A total of 326 contacts were identified and underwent three rounds of screening and one follow up examination. In the initial screening, 35 close contacts from the same class showed strong TST positivity, corresponding to a strong positivity rate of 55.56%, significantly higher than the 20.76% observed among casual contacts ( χ 2=29.80, P <0.01). Among the 35 strongly TST positivvity close contacts and five individuals with moderate TST positivity whose induration had increased by ≥10 mm over two years, none received timely preventive treatment initially; five of them were subsequently diagnosed with active TB within three months. Following this, 25 individuals initiated preventive therapy, resulting in a preventive treatment initiation rate of 62.50%. Among TST negative classmates who converted to strong positivity on repeat TST testing at three months, 75.00% started preventive treatment, but only 22.22% completed the full course. Conclusion Inadequate implementation of morning health checks and cause tracking for absenteeism due to illness, poorly standardized screening procedures, and delayed preventive treatment may have been key factors contributing to the spread of the outbreak.

[Objective]To summarize the clinical experience and the common herb pairs of Professor WANG Zhen in treating pulmonary nodules based on"resolving method".[Methods]To collect and analyse the common herb pairs,compatibility characteristics,drug characteristics and common dosage of WANG's experience in treating pulmonary nodules by copying the prescriptions;and a case was attached as evidence.[Results]Professor WANG believes that pulmonary nodules are the sthenia pathogenic factors caused by the accumulation of pathological products such as Qi stagnation,phlegm dampness,blood stasis and toxic pathogenic factors.Based on the"resolving method",he specifically adopts the methods of breaking Qi and promoting Qi,promoting blood circulation and removing blood stasis,expelling phlegm and removing dampness,softening and dispersing,and resolving stagnation and toxic factors."Sparganii Rhizoma-Curcumae Rhizoma""Citri Sarcodactylis Fructus-Aurantii Fructus""Persicae Semen-Carthami Flos""Angelicae Sinensis Radix-Lycopi Herba""Tsaoko Fructus-Atractylodis Macrocephalae Rhizoma""Prunellae Spica-Fritillariae Thunbergii Bulbus""Sargassum-Laminariae Thallus-Chloriti Lapis""Scutellariae barbatae Herba-Lobeliae chinensis Herba-Hedyotis diffusa Willd"are commonly used,the treatment is modified according to the symptoms,and the curative effect is remarkable.The patient in the case was characterized by phlegm-stasis blocking.The treatment was performed by promoting blood circulation and removing blood stasis,dispelling phlegm and dredging channel.Based on the"resolving method",the above-mentioned herb pairs were applied and modified according to the symptoms,and the curative effect was remarkable.[Conclusion]Professor WANG uses"resolving method"to treat pulmonary nodules,the selection of drugs is far-sighted and the herb pairs are concise and effective,which is worth further study.

Objective To explore the value of low contrast dose and low flow rate one-stop craniocervical CT angiography(CT A)-cerebral CT perfusion(CTP)for detecting carotid atherosclerosis(CAS).Methods Totally 117 CAS patients were prospectively enrolled and divided into group A(n=19),B(n=52),C(n=46),and low contrast dose and low flow rate one-stop craniocervical CTA-brain CTP scanning,low contrast dose and low flow rate craniocervical CT A scanning,as well as conventional craniocervical CT A scanning were performed,respectively.Virtual monoenergetic images(VMI)of 40,50 and 60 keV were reconstructed in group A and B.The subjective and objective evaluations of image quality were compared among 3 groups.Results Subjective scores of image quality and diagnostic confidence of 40 and 50 keV VMI,and the diagnostic confidence of 60 keV VMI in group A and B were not significant different compared with those in group C(all P＞0.05).Signal-to-noise ratio(SNR)and contrast-to-noise ratio(CNR)of each segment of craniocervical blood vessels at 40 and 50 keV VMI in group A and B were all higher than those in group C(all P＜0.05).CNR of cavernous sinus segment of internal carotid artery(C5 segment),horizontal segment of middle cerebral artery(MCA)(A1 segment),lateral sulcus segment of MCA(M2 segment)and basilar artery(BA)segment in group A at 60 keV VMI were all higher than those in group C(all P＜0.05).SNR of C5 segment,A1 segment and BA segment,as well as CNR of BA segment of 60 keV VMI in group B were all higher than those in group C(all P＜0.05).Conclusion Low contrast dose and low flow rate one-stop craniocervical was feasible for detecting CAS.

Objective To explore the effects of graphene quantum dots(GQDs)on the reprogramming of Müller cells.Methods Müller cells were randomly divided into the control,dedifferentiation,dedifferentiation+GQD,and GQD groups.The cells in the dedifferentiation group and the dedifferentiation+GQD group were dedifferentiated using the serum-free medium containing DMEM/F12(1∶1),20 μg·L-1 EGF,10 μg·L-1 bFGF,2 × B27,and 1 × N2 for 5 d.Then,the cells in the dedifferentiation+GQD group and the GQD group were treated with 50 mg·L-1 GQDs for 72 h.The Müller cells in the control group were subjected to normal cell culture.Immunofluorescence staining was used to identify the ex-pression of Müller cell markers,including glial fibrillary acidic protein(GFAP),glutamate-aspartate transporter(GLAST),and glutamine synthetase(GS).Phalloidin staining was performed to observe whether Müller cells could take up GQDs.The effect of different concentrations of GQDs on the proliferation of Müller cells was assessed using the cell counting kit-8(CCK-8)assay.The effect of GQDs on the expression of neural progenitor/stem cell markers(SRY-box transcription factor 2[SOX-2]and Nestin)and the astrocyte marker GFAP in normal and dedifferentiated Müller cells was examined using im-munofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluo-rescence of GFAP was extremely weak and almost invisible in Müller cells,while the fluorescence of GLAST and GS was extremely strong and predominantly appeared in the cytoplasm of Müller cells.After 24 h of GQD treatment,trace amounts of GQD fluorescence were visible in Müller cells.The amount of GQD fluorescence in the cytoplasm of Müller cells gradual-ly increased with time.The CCK-8 assay results showed that the activity of Müller cells tended to decrease with an increase in the treatment time and concentration of GQD.The statistical analysis results showed that the mean fluorescence intensity of GFAP,Nestin,and SOX-2 in Müller cells of the dedifferentiation,dedifferentiation+GQD,and GQD groups was higher than that of the control group,and the differences were statistically significant(all P＜0.05).The mean fluorescence inten-sity of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the mean fluores-cence intensity of Nestin and SOX-2 was lower than that in the dedifferentiation group,and the differences were all statisti-cally significant(P＜0.05).The relative protein expression of GFAP,Nestin,and SOX-2 in Müller cells in the dedifferentia-tion,dedifferentiation+GQD,and GQD groups was higher than that in the control group,and the differences were statis-tically significant(all P＜0.05).The relative protein expression of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the relative protein expression of Nestin and SOX-2 was lower than that in the dedi-fferentiation group,and the differences were statistically significant(all P＜0.05).Conclusion GQDs facilitate the re-programming of Müller cells into astrocytes.

Objective To explore the effects of graphene quantum dots(GQDs)on the reprogramming of Müller cells.Methods Müller cells were randomly divided into the control,dedifferentiation,dedifferentiation+GQD,and GQD groups.The cells in the dedifferentiation group and the dedifferentiation+GQD group were dedifferentiated using the serum-free medium containing DMEM/F12(1∶1),20 μg·L-1 EGF,10 μg·L-1 bFGF,2 × B27,and 1 × N2 for 5 d.Then,the cells in the dedifferentiation+GQD group and the GQD group were treated with 50 mg·L-1 GQDs for 72 h.The Müller cells in the control group were subjected to normal cell culture.Immunofluorescence staining was used to identify the ex-pression of Müller cell markers,including glial fibrillary acidic protein(GFAP),glutamate-aspartate transporter(GLAST),and glutamine synthetase(GS).Phalloidin staining was performed to observe whether Müller cells could take up GQDs.The effect of different concentrations of GQDs on the proliferation of Müller cells was assessed using the cell counting kit-8(CCK-8)assay.The effect of GQDs on the expression of neural progenitor/stem cell markers(SRY-box transcription factor 2[SOX-2]and Nestin)and the astrocyte marker GFAP in normal and dedifferentiated Müller cells was examined using im-munofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluo-rescence of GFAP was extremely weak and almost invisible in Müller cells,while the fluorescence of GLAST and GS was extremely strong and predominantly appeared in the cytoplasm of Müller cells.After 24 h of GQD treatment,trace amounts of GQD fluorescence were visible in Müller cells.The amount of GQD fluorescence in the cytoplasm of Müller cells gradual-ly increased with time.The CCK-8 assay results showed that the activity of Müller cells tended to decrease with an increase in the treatment time and concentration of GQD.The statistical analysis results showed that the mean fluorescence intensity of GFAP,Nestin,and SOX-2 in Müller cells of the dedifferentiation,dedifferentiation+GQD,and GQD groups was higher than that of the control group,and the differences were statistically significant(all P＜0.05).The mean fluorescence inten-sity of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the mean fluores-cence intensity of Nestin and SOX-2 was lower than that in the dedifferentiation group,and the differences were all statisti-cally significant(P＜0.05).The relative protein expression of GFAP,Nestin,and SOX-2 in Müller cells in the dedifferentia-tion,dedifferentiation+GQD,and GQD groups was higher than that in the control group,and the differences were statis-tically significant(all P＜0.05).The relative protein expression of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the relative protein expression of Nestin and SOX-2 was lower than that in the dedi-fferentiation group,and the differences were statistically significant(all P＜0.05).Conclusion GQDs facilitate the re-programming of Müller cells into astrocytes.

[Objective]To summarize the clinical experience and the common herb pairs of Professor WANG Zhen in treating pulmonary nodules based on"resolving method".[Methods]To collect and analyse the common herb pairs,compatibility characteristics,drug characteristics and common dosage of WANG's experience in treating pulmonary nodules by copying the prescriptions;and a case was attached as evidence.[Results]Professor WANG believes that pulmonary nodules are the sthenia pathogenic factors caused by the accumulation of pathological products such as Qi stagnation,phlegm dampness,blood stasis and toxic pathogenic factors.Based on the"resolving method",he specifically adopts the methods of breaking Qi and promoting Qi,promoting blood circulation and removing blood stasis,expelling phlegm and removing dampness,softening and dispersing,and resolving stagnation and toxic factors."Sparganii Rhizoma-Curcumae Rhizoma""Citri Sarcodactylis Fructus-Aurantii Fructus""Persicae Semen-Carthami Flos""Angelicae Sinensis Radix-Lycopi Herba""Tsaoko Fructus-Atractylodis Macrocephalae Rhizoma""Prunellae Spica-Fritillariae Thunbergii Bulbus""Sargassum-Laminariae Thallus-Chloriti Lapis""Scutellariae barbatae Herba-Lobeliae chinensis Herba-Hedyotis diffusa Willd"are commonly used,the treatment is modified according to the symptoms,and the curative effect is remarkable.The patient in the case was characterized by phlegm-stasis blocking.The treatment was performed by promoting blood circulation and removing blood stasis,dispelling phlegm and dredging channel.Based on the"resolving method",the above-mentioned herb pairs were applied and modified according to the symptoms,and the curative effect was remarkable.[Conclusion]Professor WANG uses"resolving method"to treat pulmonary nodules,the selection of drugs is far-sighted and the herb pairs are concise and effective,which is worth further study.

Objective To explore the value of low contrast dose and low flow rate one-stop craniocervical CT angiography(CT A)-cerebral CT perfusion(CTP)for detecting carotid atherosclerosis(CAS).Methods Totally 117 CAS patients were prospectively enrolled and divided into group A(n=19),B(n=52),C(n=46),and low contrast dose and low flow rate one-stop craniocervical CTA-brain CTP scanning,low contrast dose and low flow rate craniocervical CT A scanning,as well as conventional craniocervical CT A scanning were performed,respectively.Virtual monoenergetic images(VMI)of 40,50 and 60 keV were reconstructed in group A and B.The subjective and objective evaluations of image quality were compared among 3 groups.Results Subjective scores of image quality and diagnostic confidence of 40 and 50 keV VMI,and the diagnostic confidence of 60 keV VMI in group A and B were not significant different compared with those in group C(all P＞0.05).Signal-to-noise ratio(SNR)and contrast-to-noise ratio(CNR)of each segment of craniocervical blood vessels at 40 and 50 keV VMI in group A and B were all higher than those in group C(all P＜0.05).CNR of cavernous sinus segment of internal carotid artery(C5 segment),horizontal segment of middle cerebral artery(MCA)(A1 segment),lateral sulcus segment of MCA(M2 segment)and basilar artery(BA)segment in group A at 60 keV VMI were all higher than those in group C(all P＜0.05).SNR of C5 segment,A1 segment and BA segment,as well as CNR of BA segment of 60 keV VMI in group B were all higher than those in group C(all P＜0.05).Conclusion Low contrast dose and low flow rate one-stop craniocervical was feasible for detecting CAS.

OBJECTIVE To analyze the characteristics of severe cutaneous adverse reactions(SCARs)induced by oral anticoagulants(OACs),and provide a reference for clinical safety of drug use.METHODS Case reports of SCARs caused by OACs(warfarin,apixaban,rivaroxaban,edoxaban,dabigatran etexilate)were retrieved from PubMed,Embase,CNKI,Wanfang Data,VIP and other databases with search terms as"oral anticoagulants""factor Ⅹa inhibitor""direct thrombin inhibitor"and their Chinese equivalents.A descriptive statistical analysis was performed.RESULTS A total of 11 articles were included,involving 11 patients in total,among whom there were 5 males(45.5%)and 6 females(54.5%),with an average age of(59.6±21.5)years.The primary underlying diseases were mainly atrial fibrillation,pulmonary embolism,joint replacement and valve replacement.The OACs involved included warfarin in 3 cases,rivaroxaban in 4 cases,apixaban in 2 cases,and dabigatran etexilate in 2 cases.SCARs occurred from 10 hours to 42 days after treatment,and 7 cases(63.6%)within 10 to 28 days.Among 11 patients,5 cases were diagnosed as drug reaction with eosinophilia and systemic symptoms,4 cases were diagnosed as Stevens-Johnson syndrome or toxic epidermal necrolysis,and 2 cases were diagnosed as acute generalized exanthematous pustulosis.The clinical manifestations mainly included rash,fever and mucosal damage,etc.Except for 1 patient who died of sepsis and diffuse intravascular coagulation,the rest of the patients improved or recovered after withdrawal and treatment with glucocorticoids.CONCLUSIONS SCARs are rare but serious adverse reactions caused by OACs,typically occurring 10 to 28 days after medication.Once SCARs are suspected to be caused by OACs,the medication should be discontinued immediately,and a treatment plan should be formulated based on the type of SCARs to ensure the safety of patients'drug use.